LOCUS       V036913                 7759 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036913
VERSION     V036913
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3249
                     /label="PRKG2(NM_006259)"
                     /note="PRKG2(NM_006259)"
                     /gene="PRKG2"
     CDS             3256..3321
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3359..3583
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3629..4057
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4071..4400
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4467..5261
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5435..5556
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5605..5621)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5629..5645
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5653..5683)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5698..5719
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6007..6592)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6763..7623)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7624..7728)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgggaa atggttcagt gaaacctaaa cattctaagc acccagatgg acactctggg
     1021 aacctcacca ctgatgctct gcggaacaag gtgacagagc tggagagaga gttgaggagg
     1081 aaggatgctg agatccagga gcgggagtac catttgaagg agctgcggga gcagctgtcg
     1141 aagcagactg tggccattgc tgaactcaca gaggagctcc agaacaagtg catccagctg
     1201 aacaagctgc aggatgtggt gcatatgcag ggaggaagcc cgcttcaggc ctctccagat
     1261 aaagtgcctc ttgaggtcca ccggaagacc tctggattgg tctctctcca tagcaggagg
     1321 ggagcaaagg ctggcgtgtc tgctgagcca acaacccgga cctatgacct gaacaaaccc
     1381 cctgaatttt cctttgagaa agcaagagtc agaaaagact ccagtgagaa gaagctcatt
     1441 acagatgccc ttaataaaaa tcagtttctg aaaagactgg atcctcagca gatcaaagac
     1501 atggtggaat gcatgtatgg gagaaactat cagcaaggga gttacattat taagcaagga
     1561 gaaccaggaa accatatctt tgtgctggca gagggtcgac tagaggtgtt ccaaggggag
     1621 aaattgctgt cctccatccc tatgtggacc acatttgggg agcttgccat tttatacaat
     1681 tgtacaagga ctgcctctgt gaaagctatt accaatgtta aaacatgggc actagatcga
     1741 gaggtattcc agaatataat gaggaggaca gcccaagcta gagatgaaca atacagaaac
     1801 ttcctcagaa gtgtatcctt gctgaagaat ttacctgaag ataaattaac caagatcatt
     1861 gactgcttgg aagtggaata ctatgacaaa ggagattaca tcattagaga gggcgaggaa
     1921 ggaagtacct ttttcatttt ggcaaaagga aaggtaaaag taacacagag cacagaaggc
     1981 catgatcaac cacagctgat aaaaacactg cagaaaggag aatactttgg agaaaaagct
     2041 cttatcagtg atgatgtcag gtcagctaac attattgctg aagaaaatga tgttgcatgc
     2101 ctggttatag atcgagaaac attcaaccaa actgtcggta catttgaaga gctgcaaaaa
     2161 taccttgaag gatatgtggc aaacctgaac cgtgatgatg aaaaaagaca tgcgaagcgg
     2221 tccatgtcta actggaagct gtccaaagca ctctctctgg aaatgattca gctgaaggag
     2281 aaggtggcca gattttcctc atcatcccca ttccagaacc ttgagattat tgcaacactg
     2341 ggcgttggtg ggttcggaag agttgagctt gttaaagtaa aaaatgagaa tgttgctttt
     2401 gctatgaagt gtataaggaa gaagcacata gttgacacca agcagcagga gcatgtctac
     2461 tcagagaaga ggatcctaga ggagctgtgc tctccattca ttgtgaaatt atatcgtact
     2521 ttcaaggaca ataagtatgt atacatgctt ctggaggcct gcttaggtgg ggagctctgg
     2581 agtatattaa gggacagagg cagctttgat gaacccacct ccaaattctg cgttgcttgt
     2641 gtgacagaag catttgatta cctgcatcga ctaggtatta tctacagaga cttgaaacca
     2701 gaaaacttaa ttctagatgc tgagggttac cttaaattgg ttgactttgg atttgcgaag
     2761 aaaatagggt ctggacagaa aacatggaca ttctgtggga ctccagaata tgtagctcct
     2821 gaagtcattc tcaacaaggg acatgacttc agtgtggatt tctggtcact gggaattcta
     2881 gtgtatgagc tcctaacggg caacccaccc ttttctgggg ttgaccaaat gatgacctac
     2941 aatttgattc tcaaaggaat tgaaaaaatg gattttccca ggaagataac acgacgacct
     3001 gaggatttga ttcggaggct ttgcaggcaa aatccaacag aaaggctggg aaatctgaag
     3061 aatggaataa atgacattaa gaaacacagg tggttaaatg gttttaattg ggagggactg
     3121 aaagcacgga gccttccatc acctttgcaa agagagctca agggacccat agatcacagc
     3181 tactttgaca aatatcctcc tgaaaaggga atgcctccag atgagctatc aggctgggat
     3241 aaagacttcc tcgaggacta caaagaccat gacggtgatt ataaagatca tgacatcgac
     3301 tacaaggatg acgatgacaa gtagtgaggg cccgtttaaa cccgctgatc agcctcgact
     3361 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3421 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3481 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3541 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3601 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3661 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3721 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3781 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3841 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3901 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3961 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4021 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4081 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4141 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4201 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4261 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4321 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4381 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4441 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4501 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4561 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4621 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4681 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4741 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4801 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4861 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4921 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4981 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5041 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5101 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5161 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5221 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5281 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5341 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5401 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5461 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5521 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5581 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5641 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5701 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5761 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5821 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5881 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5941 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6001 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6061 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6121 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6181 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6241 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6301 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6361 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6421 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6481 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6541 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6601 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6661 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6721 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6781 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6841 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6901 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6961 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7021 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7081 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7141 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7201 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7261 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7321 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7381 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7441 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7501 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7561 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7621 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7681 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7741 aaaagtgcca cctgacgtc
//
